[Detection of trehalose in transgenic tobacco by HPLC with ELSD]

The E. coli trehase synthalose gene(otsA) was transferred into Nicotiana tabacum mediated by Agrobacterium, but the method for detecting low concentration of trehalose in transgenic plant was not available. The high performance liquid chromatograph(HPLC) with evaporative light-scatting detector(ELSD) using water: methyl cyanide(1:2.6 v/v) as mobile phase was established in this work. An ODS column Zorbax RX-SIL was employed. the trehalose detection limits of ELSD was 5 mg/L. From the linear relationship between the logarithm of trehalose concentration and the logarithm of peak area, it was shown there was 14.7 micrograms.(g FW)-1 in transgenic plant. The data strongly confirmed that trehalose was responsible for the improved stress tolerance of the tobacco.